The Preparation and Identification of P2X7 Short Peptide ReceptorMonoclonal Antibody

Biotechnology Bulletin ›› 2012, Vol. 0 ›› Issue (12): 167-172.

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The Preparation and Identification of P2X7 Short Peptide ReceptorMonoclonal Antibody

Tan Chao^1,2, Han Li¹, Shen Xiejing¹

1 Medical Science College of China Three Gorges University，Yichang 443002 ；2Department of Internal Medicine Yichang Central People’sHospital Laboratory Medicine，the First Clinical Medical College of China Three Gorges University，Yichang 443002

Received:2012-07-06 Revised:2013-01-25 Online:2012-12-26 Published:2013-02-06

Abstract

Abstract: The aim was to prepare and identify the monoclonal antibodies of the P2X7 receptor. Our method was to use the extracellularfragment of human beings’ P2X7 receptor to produce the short peptide as an antigen to immunize mouse for three times. Then B lymphocyteswere isolated from the mouse spleen, and then mixed with myeloma cells and cultured. The positive hybridoma clones were selected and widelycultured to prepare and identify the biological effects of P2X7. The supernatant was collected and purified by HiTrap protein G affinity column.The results were as follows. Firstly, a stable hybridoma cell line secreting mAb against human P2X7 receptor was obtained, and the heavy andlight chains of the secreted mAb were IgG1 and κ respectively. The titers of the ascites was 1∶6.4×104, which was remained stable aftersubculture of hybridoma cells for 30 passages or storage at liquid nitrogen for 6 months. Secondly, the western blotting test showed that the mAbcould specifically bind to P2X7 receptor. The conclusion was that the prepared mAb against P2X7 receptor showed high specificity and stability,which laid a foundation for the development of the antibody drug aimed at P2X7 receptor as a target antigen.

Tan Chao, Han Li, Shen Xiejing. The Preparation and Identification of P2X7 Short Peptide ReceptorMonoclonal Antibody[J]. Biotechnology Bulletin, 2012, 0(12): 167-172.

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The Preparation and Identification of P2X7 Short Peptide ReceptorMonoclonal Antibody

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